Cargando…
Metabolic engineering of Rhodotorula toruloides for resveratrol production
BACKGROUND: Resveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial production of re...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9789595/ https://www.ncbi.nlm.nih.gov/pubmed/36566171 http://dx.doi.org/10.1186/s12934-022-02006-w |
_version_ | 1784858989375258624 |
---|---|
author | Zhang, Mengyao Gao, Qidou Liu, Yijuan Fang, Zhumei Gong, Zhiwei Zhao, Zongbao K. Yang, Xiaobing |
author_facet | Zhang, Mengyao Gao, Qidou Liu, Yijuan Fang, Zhumei Gong, Zhiwei Zhao, Zongbao K. Yang, Xiaobing |
author_sort | Zhang, Mengyao |
collection | PubMed |
description | BACKGROUND: Resveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial production of resveratrol offers a promising alternative to plant- and chemical-based processes. The non-conventional oleaginous yeast Rhodotorula toruloides is a potential workhorse for the production of resveratrol that endowed with an efficient and intrinsic bifunctional phenylalanine/tyrosine ammonia-lyase (RtPAL) and malonyl-CoA pool, which may facilitate the resveratrol synthesis when properly rewired. RESULTS: Resveratrol showed substantial stability and would not affect the R. toruloides growth during the yeast cultivation in flasks. The heterologus resveratrol biosynthesis pathway was established by introducing the 4-coumaroyl-CoA ligase (At4CL), and the stilbene synthase (VlSTS) from Arabidopsis thaliana and Vitis labrusca, respectively. Next, The resveratrol production was increased by 634% through employing the cinnamate-4-hydroxylase from A. thaliana (AtC4H), the fused protein At4CL::VlSTS, the cytochrome P450 reductase 2 from A. thaliana (AtATR2) and the endogenous cytochrome B5 of R. toruloides (RtCYB5). Then, the related endogenous pathways were optimized to affect a further 60% increase. Finally, the engineered strain produced a maximum titer of 125.2 mg/L resveratrol in YPD medium. CONCLUSION: The non-conventional oleaginous yeast R. toruloides was engineered for the first time to produce resveratrol. Protein fusion, co-factor channeling, and ARO4 and ARO7 overexpression were efficient for improving resveratrol production. The results demonstrated the potential of R. toruloides for resveratrol and other phenylpropanoids production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-02006-w. |
format | Online Article Text |
id | pubmed-9789595 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-97895952022-12-25 Metabolic engineering of Rhodotorula toruloides for resveratrol production Zhang, Mengyao Gao, Qidou Liu, Yijuan Fang, Zhumei Gong, Zhiwei Zhao, Zongbao K. Yang, Xiaobing Microb Cell Fact Research BACKGROUND: Resveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial production of resveratrol offers a promising alternative to plant- and chemical-based processes. The non-conventional oleaginous yeast Rhodotorula toruloides is a potential workhorse for the production of resveratrol that endowed with an efficient and intrinsic bifunctional phenylalanine/tyrosine ammonia-lyase (RtPAL) and malonyl-CoA pool, which may facilitate the resveratrol synthesis when properly rewired. RESULTS: Resveratrol showed substantial stability and would not affect the R. toruloides growth during the yeast cultivation in flasks. The heterologus resveratrol biosynthesis pathway was established by introducing the 4-coumaroyl-CoA ligase (At4CL), and the stilbene synthase (VlSTS) from Arabidopsis thaliana and Vitis labrusca, respectively. Next, The resveratrol production was increased by 634% through employing the cinnamate-4-hydroxylase from A. thaliana (AtC4H), the fused protein At4CL::VlSTS, the cytochrome P450 reductase 2 from A. thaliana (AtATR2) and the endogenous cytochrome B5 of R. toruloides (RtCYB5). Then, the related endogenous pathways were optimized to affect a further 60% increase. Finally, the engineered strain produced a maximum titer of 125.2 mg/L resveratrol in YPD medium. CONCLUSION: The non-conventional oleaginous yeast R. toruloides was engineered for the first time to produce resveratrol. Protein fusion, co-factor channeling, and ARO4 and ARO7 overexpression were efficient for improving resveratrol production. The results demonstrated the potential of R. toruloides for resveratrol and other phenylpropanoids production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-02006-w. BioMed Central 2022-12-24 /pmc/articles/PMC9789595/ /pubmed/36566171 http://dx.doi.org/10.1186/s12934-022-02006-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Zhang, Mengyao Gao, Qidou Liu, Yijuan Fang, Zhumei Gong, Zhiwei Zhao, Zongbao K. Yang, Xiaobing Metabolic engineering of Rhodotorula toruloides for resveratrol production |
title | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
title_full | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
title_fullStr | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
title_full_unstemmed | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
title_short | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
title_sort | metabolic engineering of rhodotorula toruloides for resveratrol production |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9789595/ https://www.ncbi.nlm.nih.gov/pubmed/36566171 http://dx.doi.org/10.1186/s12934-022-02006-w |
work_keys_str_mv | AT zhangmengyao metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction AT gaoqidou metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction AT liuyijuan metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction AT fangzhumei metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction AT gongzhiwei metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction AT zhaozongbaok metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction AT yangxiaobing metabolicengineeringofrhodotorulatoruloidesforresveratrolproduction |